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. 2015 Dec 21;10(12):e0145336. doi: 10.1371/journal.pone.0145336

Fig 4. The HMG domain of Sox2 mediates its direct binding to Ash2l of Set1/Mll complexes.

Fig 4

(A) Diagram for truncation mutants of FLAG-HA-tagged Sox2 showing their domain structures and whether the protein binds to Ash2l as summarized from (B). (B) 293T cells were transfected with indicated FLAG-HA-tagged Sox2 mutants, and co-IP was performed by anti-FLAG M2 resin. Wdr5 blotting for the right panel is not shown due to a fortuitous cross-reactivity with ΔHMG. TAD: transcriptional activation domain. Inp, input (2.5%); F↓, anti-FLAG immunoprecipitation. (C) Purified proteins were examined by SDS-PAGE along with the molecular weight ladder on the left, and visualized by coomassie blue staining. (D) In vitro binding assay for Ni bead-bound His-Sox2 with FLAG-Ash2l and FLAG-Wdr5, and examined by western blotting using anti-FLAG antibody. Ni-beads that was not used for His-Sox2 purification was used as the mock control in the binding assay. Input (20%) was loaded.